Kespohl Sabine, Jost Robin, Maryska Silke, Altin Lena-Maria, Sander Ingrid, Schülke Stefan, Paulus-Tremel Kathrin E, Bonertz Andreas, Klose Thomas, Mahler Vera, Raulf Monika
Institute for Prevention and Occupational Medicine of the DGUV, Ruhr-University Bochum (IPA), Bochum.
Division Allergology, Paul-Ehrlich-Institut, Langen, and.
Allergol Select. 2024 Jul 4;8:238-250. doi: 10.5414/ALX02506E. eCollection 2024.
In order to ensure valid diagnostics for occupational test allergen solutions despite the ongoing reduction in the availability of commercial test extracts, a plan B was initiated for the possible production of skin prick test (SPT) solutions in public pharmacies. For important occupational allergen sources (wheat and rye, storage mites, animal epithelia, mold material) laboratory extraction methods were analyzed in comparison to pharmacy compatible extraction methods regarding protein quantity and quality in SDS-PAGE combined with silver staining. Subsequently, using the example of bovine epithelia, adapted extraction procedures as well as in-process and final product controls were transferred to a public pharmacy. Allergen sources with a high protein content, such as wheat and rye grains as well as storage mites, showed good comparability of the extractable protein quantity and protein pattern, regardless of the applied extraction method. In contrast, allergen source materials with a low total protein content, such as animal epithelia and molds, can benefit from laboratory extraction conditions such as mechanical disruption and specific buffer additives. In the qualitative protein silver staining, characteristic protein patterns were identified for each allergen source. Depending on the extraction method, only minor differences in total protein patterns were observed in animal epithelia and molds. Using source materials from two suppliers, the resulting allergen extracts displayed clear differences in protein content in storage mites and quantitative and qualitative differences in molds. A practical preparation attempt of SPT solutions in a public pharmacy was successful. SPT solutions prepared with adapted pharmacy extraction methods showed a comparable protein and Bos d 2 allergen content and equivalent qualities in the protein pattern compared to a previously available commercial SPT solution. Accordingly, it can be assumed that standardized SPT solutions with sufficient allergen quality for occupational allergen sources can be prepared in public pharmacies if certified allergen sources with appropriate protein content are available.
为了在商业测试提取物的可获得性持续减少的情况下确保对职业测试变应原溶液进行有效的诊断，启动了B计划，以便在公共药房中可能生产皮肤点刺试验（SPT）溶液。对于重要的职业变应原来源（小麦和黑麦、仓储螨、动物上皮、霉菌材料），将实验室提取方法与药房兼容提取方法在SDS - 聚丙烯酰胺凝胶电泳（SDS - PAGE）结合银染法中的蛋白质数量和质量方面进行了分析比较。随后，以牛上皮为例，将经过调整的提取程序以及过程中和最终产品控制方法转移到了一家公共药房。蛋白质含量高的变应原来源，如小麦和黑麦谷物以及仓储螨，无论采用何种提取方法，其可提取蛋白质数量和蛋白质图谱都具有良好的可比性。相比之下，总蛋白质含量低的变应原来源材料，如动物上皮和霉菌，可受益于实验室提取条件，如机械破碎和特定的缓冲添加剂。在蛋白质银染定性分析中，确定了每种变应原来源的特征性蛋白质图谱。根据提取方法的不同，在动物上皮和霉菌中仅观察到总蛋白质图谱的微小差异。使用来自两个供应商的原料，所得变应原提取物在仓储螨的蛋白质含量以及霉菌的定量和定性方面显示出明显差异。在一家公共药房进行的SPT溶液实际制备尝试取得了成功。与先前可用的商业SPT溶液相比，采用经过调整的药房提取方法制备的SPT溶液在蛋白质和牛Bos d 2变应原含量以及蛋白质图谱质量方面具有可比性。因此，可以假设，如果有蛋白质含量合适的经过认证的变应原来源，在公共药房中可以制备出具有足够变应原质量用于职业变应原来源的标准化SPT溶液。
